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Fusion protein, corresponding to amino acids 1759-2093 of Human Mitosin/CENPF.
Clone 1D8 recognizes mitosin, a 350-kDa phosphoprotein, that is specifically involved in mitotic-phase progression. Mitosin is expressed throughout S, G2, and M phases of the cell cycle but is absent in G0 and G1.
This antibody could be of use as a proliferation marker. This protein associates with the centromere-kinetochore complex. The protein is a component of the nuclear matrix during the G2 phase of interphase. In late G2 the protein associates with the kinetochore and maintains this association through early anaphase. It localizes to the spindle midzone and the intracellular bridge in late anaphase and telophase, respectively, and is thought to be subsequently degraded. The localization of this protein suggests that it may play a role in chromosome segregation during mitotis. It is thought to form either a homodimer or heterodimer. Autoantibodies against this protein have been found in patients with cancer or graft versus host disease.
Our Abpromise guarantee covers the use of ab90 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 367 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|Flow Cyt||Use 0.5µg for 106 cells.|
CENPF was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to CENPF (ab90)and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab90. Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Bands: 370kDa:CENPF; 171kDa: non specific - as present in control (lane 2); 70kDa: We are unsure as to the identity of this extra band.